Difference between revisions of "Cell and tissue lysis hub"

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(Comparison of lysis methods)
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[[Image:Hub icon.png|right|thumbnail|'''protocol hub''']]
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[[Image:Hub icon.png|right]]
  
This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison.
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This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison. The ups and downs of various methods from sonication, homogenization, freeze-thaw cycles, and detergent-based lysis are examined below.
  
 
== Comparison of lysis methods ==
 
== Comparison of lysis methods ==
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* problem: heat build up which can denature proteins (proportionate to length of sonication)
 
* problem: heat build up which can denature proteins (proportionate to length of sonication)
 
* precaution: do on ice and sonicate intermittantly
 
* precaution: do on ice and sonicate intermittantly
 +
material: ultrasonication bath or rods
  
 
=== Homogenisation ===  
 
=== Homogenisation ===  
 
* best for animal tissue; less suitable for cells
 
* best for animal tissue; less suitable for cells
 
* precaution: do on ice to reduce heat build-up and denaturation
 
* precaution: do on ice to reduce heat build-up and denaturation
 +
material: drills (polytron), pestle/tube (dounce) bead beaters
  
 
=== Freeze-thaw ===
 
=== Freeze-thaw ===
 
* least effective method
 
* least effective method
 
* plus: does not denature proteins as much as other methods
 
* plus: does not denature proteins as much as other methods
 +
material: pestle and mortar, liquid nitrogen
  
 
=== Detergents ===
 
=== Detergents ===
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* problems: detergent may inhibit subsequent reactions
 
* problems: detergent may inhibit subsequent reactions
 
* problems: detergent may disrupts protein interactions
 
* problems: detergent may disrupts protein interactions
 +
no additional material required, just the chemicals and typical tubes
  
 
== Specific protocols ==
 
== Specific protocols ==
  
{| {{table}}
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{| {{sorttable}}
 
! style="background:lightgrey"|description/link
 
! style="background:lightgrey"|description/link
 
! style="background:lightgrey"|target
 
! style="background:lightgrey"|target
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| protein
 
| protein
 
| detergent (Triton)
 
| detergent (Triton)
| eukaryotic cells
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| cells
 
|-
 
|-
 
| [[Sauer:Lysing E. coli with Lysozymes]]
 
| [[Sauer:Lysing E. coli with Lysozymes]]
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| protein
 
| protein
 
| detergent (RIPA buffer)
 
| detergent (RIPA buffer)
| cell line
+
| cells
 
|-
 
|-
 
| [[Streptomyces:Protocols/Mini-Maxi Prep]]
 
| [[Streptomyces:Protocols/Mini-Maxi Prep]]
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| protein
 
| protein
 
| detergent (SDS, NP40)
 
| detergent (SDS, NP40)
| cell line
+
| cells
 
|-
 
|-
 
| [[Eccles:Protein Lysates from Tissue]]
 
| [[Eccles:Protein Lysates from Tissue]]
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| RNA
 
| RNA
 
| phenol
 
| phenol
| cell line
+
| cells
 +
|-
 +
| [[Sauer:RNA Purification from E. coli]]
 +
| RNA
 +
| various options
 +
| bacteria
 
|-
 
|-
 
| add another method's name
 
| add another method's name

Revision as of 02:45, 19 December 2008

Hub icon.png

This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison. The ups and downs of various methods from sonication, homogenization, freeze-thaw cycles, and detergent-based lysis are examined below.

Comparison of lysis methods

Sonication

  • most efficient method of cell fractionation
  • problem: heat build up which can denature proteins (proportionate to length of sonication)
  • precaution: do on ice and sonicate intermittantly

material: ultrasonication bath or rods

Homogenisation

  • best for animal tissue; less suitable for cells
  • precaution: do on ice to reduce heat build-up and denaturation

material: drills (polytron), pestle/tube (dounce) bead beaters

Freeze-thaw

  • least effective method
  • plus: does not denature proteins as much as other methods

material: pestle and mortar, liquid nitrogen

Detergents

  • chemical method of lysis
  • problems: detergent may inhibit subsequent reactions
  • problems: detergent may disrupts protein interactions

no additional material required, just the chemicals and typical tubes

Specific protocols

description/link target lysis method type of material
Silver: Lysate for Western protein detergent (Triton) cells
Sauer:Lysing E. coli with Lysozymes lysozyme bacteria
Blackburn:Yeast Colony PCR DNA NaOH yeast
Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer protein detergent (RIPA buffer) cells
Streptomyces:Protocols/Mini-Maxi Prep DNA NaOH bacteria
Western Blot/Tissue Preparation protein detergent & physical disruption tissue
Eccles:Protein Lysates from Cells in Culture protein detergent (SDS, NP40) cells
Eccles:Protein Lysates from Tissue protein detergent, physical, freezing tissue
RNA extraction using trizol/tri RNA phenol cells
Sauer:RNA Purification from E. coli RNA various options bacteria
add another method's name target lysis method type of starting material

Related hub pages

Related discussions on BioForum